We are continuing our efforts to immunolocalize proteins important for cytoskeletal function in both mammalian cells and in yeast by applying antibodies to thin sections of samples that have been well-fixed by cryopreservation and freeze-substitution and embedded in the Lowicryls or LR-White. Of four antibodies recently obtained from collaborators, two have yielded well-controlled localizations with signals that are above background by statistically rigorous tests. Thus, otherwise excellent antibodies provided by Ahn against active forms of MAP kinase and by Pillus against the telomere-binding protein RAP1 have failed to give good localizations, while two antibodies from Gould have proven successful, yielding one published paper and one manuscript in preparation. Each of these papers describes some new aspect of the relationship between actin and actin-related proteins (ARPs) in fission yeast. We have also had some success with myc-tagged versions of protein, so long as there are enough tags present. One and three mycs have previously been unsuccessful, but with McCollum, we have now been successful using 13 mycs on a gene called sid2+, which is important in specifying the position of the cleavage furrow in fission yeasts, leading to another paper that is now being readied for submission. C10
| Giddings Jr, Thomas H; Morphew, Mary K; McIntosh, J Richard (2017) Preparing Fission Yeast for Electron Microscopy. Cold Spring Harb Protoc 2017: |
| Zhao, Xiaowei; Schwartz, Cindi L; Pierson, Jason et al. (2017) Three-Dimensional Structure of the Ultraoligotrophic Marine Bacterium ""Candidatus Pelagibacter ubique"". Appl Environ Microbiol 83: |
| Brown, Joanna R; Schwartz, Cindi L; Heumann, John M et al. (2016) A detailed look at the cytoskeletal architecture of the Giardia lamblia ventral disc. J Struct Biol 194:38-48 |
| Saheki, Yasunori; Bian, Xin; Schauder, Curtis M et al. (2016) Control of plasma membrane lipid homeostasis by the extended synaptotagmins. Nat Cell Biol 18:504-15 |
| Höög, Johanna L; Lacomble, Sylvain; Bouchet-Marquis, Cedric et al. (2016) 3D Architecture of the Trypanosoma brucei Flagella Connector, a Mobile Transmembrane Junction. PLoS Negl Trop Dis 10:e0004312 |
| Park, J Genevieve; Palmer, Amy E (2015) Properties and use of genetically encoded FRET sensors for cytosolic and organellar Ca2+ measurements. Cold Spring Harb Protoc 2015:pdb.top066043 |
| McCoy, Kelsey M; Tubman, Emily S; Claas, Allison et al. (2015) Physical limits on kinesin-5-mediated chromosome congression in the smallest mitotic spindles. Mol Biol Cell 26:3999-4014 |
| Höög, Johanna L; Lötvall, Jan (2015) Diversity of extracellular vesicles in human ejaculates revealed by cryo-electron microscopy. J Extracell Vesicles 4:28680 |
| Marc, Robert E; Anderson, James R; Jones, Bryan W et al. (2014) The AII amacrine cell connectome: a dense network hub. Front Neural Circuits 8:104 |
| Weber, Britta; Tranfield, Erin M; Höög, Johanna L et al. (2014) Automated stitching of microtubule centerlines across serial electron tomograms. PLoS One 9:e113222 |
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